Articles
OVULE CULTURE OF HELLEBORUS SPECIES
Article number
855_28
Pages
195 – 200
Language
English
Abstract
In order to establish protocols for embryo rescue techniques, in a first step this study aimed at the identification of culture conditions for ovules from intraspecific crosses.
The effects of dissection date after pollination and temperature during ovule culture were examined in intraspecific crossings of Helleborus niger, H. argutifolius, H. × hybridus and H. foetidus.
Ovaries were harvested 3-6 weeks after pollination, surface disinfected in 70% ethanol for 30 s, 2% sodium hypochlorite with one drop Tween for 10 min and rinsed in sterilised water three times.
Ovules were dissected from ovaries and cultured on medium based on MS solidified with 0.4% Gelrite at a pH of 5.8. Two media supplemented with 2.5 or 5% sucrose were compared.
Ovules were cultured in darkness at 24±1 or 16±1°C for 12 weeks.
Thereafter, ovules of each temperature treatment were split and one half was incubated at 6±1°C for 11 weeks, while the other half remained in the initial temperature.
Afterwards the ovules were placed back to their initial temperature.
On average of all species, 2% of the ovules dissected 3 weeks and 1% dissected 4 weeks after pollination germinated, while preparation after 5 weeks resulted in 4% and after 6 weeks in 8% germination, respectively.
The intermediate cold treatment with 6°C turned out to be very beneficial for later germination.
Furthermore, no effect of the media supplemented with either 2.5 or 5% sucrose was observed.
The effects of dissection date after pollination and temperature during ovule culture were examined in intraspecific crossings of Helleborus niger, H. argutifolius, H. × hybridus and H. foetidus.
Ovaries were harvested 3-6 weeks after pollination, surface disinfected in 70% ethanol for 30 s, 2% sodium hypochlorite with one drop Tween for 10 min and rinsed in sterilised water three times.
Ovules were dissected from ovaries and cultured on medium based on MS solidified with 0.4% Gelrite at a pH of 5.8. Two media supplemented with 2.5 or 5% sucrose were compared.
Ovules were cultured in darkness at 24±1 or 16±1°C for 12 weeks.
Thereafter, ovules of each temperature treatment were split and one half was incubated at 6±1°C for 11 weeks, while the other half remained in the initial temperature.
Afterwards the ovules were placed back to their initial temperature.
On average of all species, 2% of the ovules dissected 3 weeks and 1% dissected 4 weeks after pollination germinated, while preparation after 5 weeks resulted in 4% and after 6 weeks in 8% germination, respectively.
The intermediate cold treatment with 6°C turned out to be very beneficial for later germination.
Furthermore, no effect of the media supplemented with either 2.5 or 5% sucrose was observed.
Publication
Authors
J. Meiners, T. Winkelmann
Keywords
Helleborus, ovule culture, germination
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